Production of organic

Acids by fermentation
GROUP 4
RUTENDO SVOSVE R1811791
VIMBAINASHE MASVONGO R187519Y
BLESSING R MURIDZI R188857K
LENIN MTEMBO
NOZIPHO MAISWA R1810103
TRISH CHAKANYUKA R189860A
Citric acid
• It is a colorless weak organic acid. It occurs naturally in
citrus fruits.
• which occurs in the metabolism of all aerobic organisms.
Applications of citric acid
• Beverages- provides tartness (bitterness), Increases the
effectiveness of antimicrobial preservatives. Used in pH
adjustment to provide uniform acidity.
• Candy, Jellies, Jams- Provides tartness, acts as acidulant.
• Pharmaceuticals acts as effervescent in powders and
tablets in combination with bicarbonates. Provides rapid
dissolution of active ingredients. Acidulant in mild astringent
formulation. Anticoagulant.
MICRO-ORGANISMS USED FOR CITRIC ACID PRODUCTION

• Fungi :Aspergillus niger, Penicillium janthinelum

• Yeasts: Saccahromicopsis lipolytica
• Bacteria: Arthrobacter paraffinens
• Most of them are not able to produce commercially
acceptable yields.
• Hence, the fungus A. niger has remained the organism
of choice for commercial production_because:
i. its ease of handling,
ii. its ability to ferment a variety of cheap raw materials
iii. high yields.
PRODUCTION TECHNIQUES AND RAW MATERIALS

Submerged fermentation(SmF):
• for citric acid production the SmF process is the
commonly used technique.
• Fermenters for citric acid production require provision
of aeration system, which can maintain a high
dissolved oxygen level
• different kinds of media can be used e.g sugar and
starch based media.
• Inoculation is either by adding a suspension of spores,
or of pre-cultivated mycelia.
Fermentation continued
• when spores are used, a surfactant is added in order to
disperse them in the medium.
• For pre-cultivated mycelia, an inoculum size of 10% of
fresh medium is required.
• Normally, submerged fermentation is concluded in 5 to
10 days depending on the process conditions.
• It can be carried out in batch, continuous or fed batch
systems, although the batch mode is more frequently
used.
FACTORS AFFECTING CITRIC ACID PRODUCTION

Carbon source: Citric acid accumulation is strongly affected by the nature

of the carbon source. The presence of easily metabolized carbohydrates has
been found essential for good production of citric acid. Sucrose being the
most favourable followed by glucose, fructose
• Other sources of carbon such as ethanol, cellulose, manitol, lactic, allow a
limited growth and low production.
• A.niger strains needed an initial sugar concentration of 10-14% as
optimal; no citric acid was produced at sugar concentration of less than
2.5%.
Nitrogen source: Citric acid production is directly influenced by the nitrogen
source.
• The concentration of nitrogen source required for citric acid fermentation
is 0.1 to 0.4 N /liter.
• A high nitrogen concentration increases fungal growth and the
Factors continued
Phosphorous source: Presence of phosphate in the medium has
a great effect on the yield of citric acid.
• Potassium dihydrogen phosphate is the most suitable
phosphorous source. Low levels of phosphate favour citric acid
production,0.5 to 5.0 g/L is required by the fungus in a chemically
defined medium for maximum production of citric acid.
Trace elements:
• A number of divalent metals such as zinc, manganese, iron,
copper and magnesium have been found to affect citric acid
production by A. niger. E.g
• Citric acid accumulation decreased by the addition of iron,
• Magnesium is required both for growth as well as for citric acid
production.
Process parameters
• pH: a pH below 2.0 is required for optimum production of citric
acid. A low initial pH has the advantage of checking
contamination and inhibiting oxalic acid formation.

• Aeration: Aeration has been shown to have a determinant

effect on citric acid fermentation.
• Increased aeration rates led to enhanced yields and reduced
fermentation time
• It is important to maintain the oxygen concentration above 25%
saturation.
however high aeration rates lead to high amounts of foam,
especially during the growth phase, thus Therefore, antifoaming
agents should be added and the construction of mechanical
“defoamers” are required to tackle this problem.
Product recovery
• The recovery of citric acid from liquid fermentation is generally
accomplished by three basic procedures, precipitation, extraction, and
adsorption and absorption (mainly using ion exchange resins).
• Precipitation is performed by the addition of calcium oxide hydrate (milk
of lime) to form the slightly soluble tri-calcium citrate tetrahydrate.
• The precipitated tri-calcium citrate is removed by filtration and washed
several times with water.
• It is then treated with sulphuric acid forming calcium sulphate, which is
filtered off.
• Mother liquor containing citric acid is treated with active carbon and
passed through cation and anion exchangers.
• Finally, the liquor is concentrated in vacuum crystallizers at 20-25°C,
forming citric acid monohydrate.
• Crystalization at temperatures higher to this is used to prepare
anhydrous citric acid.
A diagram showing synthesis of Citrate using
A.niger
ITACONIC ACID
• Itaconic acid is an organic acid that is a white solid and soluble in water,
ethanol and acetone.
• Historically itaconic acid was obtained by the distillation of citric acid but
currently it is produced by fermentation.

Application
• Industrial production of resins
• Used as a building block for acrylic plastics, acrylate latexes, super
absorbents and anti scaling agents.
• Pharmaceuticals and detergents
Production
• Fermentation of sugars such as glucose or molasses.
• Using fungi Aspergillus terreus or A. itaconious from simple sugars
• Glucose is broken down into pyruvate by glycolysis which is further
converted into acetyl-CoA with the release of CO2
• Acetyl-CoA then joins the citric acid cycle where the first step from cis-
aconitate.
• Itaconic acid is also produced in cells of macrophage lineage
Lactic acid
• Lactic acid is an organic acid.
• It has a molecular formula CH₃CHCOOH.
• It is white in the solid state and it is miscible with water.
• When in the dissolved state, it forms a colourless solution
Role of Lactic acid
• Added to some processed foods to prevent spoilage and increase flavour.
• Lactic acid and the bacteria that produce it are associated with several health benefits including
improved gut health and increased nutrient absorption.
• It assists in cell respiration, glucose production and molecule signalling.
• Lactic acid build-up after a hard workout aids muscle regeneration- acts as a signal to your body that
your muscles have worked really hard and need to be repaired and rebuilt.
How is Lactic acid formed
• Lactic acid is mainly produced in muscle cells and red blood cells.
• It forms when the body breaks down carbohydrates to use for energy
when oxygen levels are low.
• Industrially, lactic acid can be produced by chemical synthesis or by
fermentation.
• Lactic acid can be produced from sugar plant wastes (molasses and sugar
beet juice), starchy and lignocellulosic biomasses.
How is Lactic acid purified
• Lactic acid is separated out and purified by using various techniques such
as precipitation by calcium hydroxide, dialysis, membrane separation,
distillation, electrodailysis, reverse osmosis, nanofiltration and ion-
exchange.
• Lactic acid process in a classical way involves a series of downstream
treatments such as precipitation, conventional filtration, acidification,
carbon adsorption, evaporation, crystallization, and others (Pal et al.
2009).
GLUCONIC ACID
• Gluconic acid is an organic compound with molecular
formula C6 H12 O7.
• In aqueous solution at neutral PH ,gluconic acid forms the
gluconate ion.
• Gluconic acid occurs naturally in honey ,wine and fruit

Applications of gluconic acid

• Gluconic acid is used in the manufacturing of food and
metal
Production of Gluconic acid
• Gluconic acid is produced from glucose ,this reaction is
catalyzed by enzyme glucose oxidase.
• Gluconic acid production by A .niger is an aerobic
fermentation with high oxygen demand.
Malic acid
• Malic acid is a white, odorless, crystalline solid. Naturally
can be extracted from fruits such as apples, litchis and
plums efforts are being made to manufacture it
industrially.
Uses
• It used in the pharmaceutical industry as compound of the
migraine drug and as a food additive (used to mask the
aftertaste of artificial sweeteners )
Production of malic acid
• It is industrially produced by the enzymatic conversion of
fumaric acid using immobilized cells of Brevibacterium
flavum.
• Malic acid can be produced by hydration of fumarate
using fumarase an enzyme of the citric cycle which
catalyses the conversion of fumaric acid to malic acid.
• The use of an immobilized biocatalyst and a continuous
flow reactor greatly improve catalyst stability and
production output
Fumaric acid
• Fumaric acid is an organic acid found widely in nature. It
is a colorless, odorless, crystalline powder with tart-fruity
taste.
• Natural sources of fumaric acid include bolete
mushrooms, lichen, and Iceland moss. It is also found in
papayas, pears and plums in very limited amounts.
• It was first isolated from the plant Fumaria officinalis,
which is where it derives its name.
• Fumaric acid is the trans-isomer of symmetric,
unsaturated dicarboxylic acid; the cis-isomer is maleic
acid.
Uses of fumaric acid
• Being the least expensive food acidulant, it is used in
beverages, baked goods, confections and dessert mixes
as a:
• Long-lasting and persistent sourness
• Substitute for tartaric and citric acids
• Acidity regulator
• Antibacterial agent and shelf-life extender
• fumaric acid is a pharmaceutically active substance that is
used to treat psoriasis or multiple sclerosis.
 Production of fumaric acid
• Fumaric acid is primarily produced using isomerization or
biochemical routes.
• Isomerization: this method is based on the conversion of
maleic anhydride to maleic acid and the cis–trans
isomerization of maleic acid to the final product.
• Biochemical: relies on bioconversion of substrates such
as corn starch, corn straw, cassava bagasse or potato
flour using maleate isomerase produced by
Pseudomonas and Arthrobacter species.
Production by fermentation
• Filamentous fungal fermentation with Rhizopus arrhizus
can produce fumaric acid from glucose via a reductive
tricarboxylic acid (TCA) pathway and was once used in
the industry before the rising of the petrochemical
industry.
• Similar to the other fungal acid fermentation processes,
the culture conditions are crucial to obtain the correct
cellular morphology, which promotes efficient production.